te70x semi-dry blotters · 2012. 10. 17. · tions de livraison n’est valable que si le produit a...

user manual

um TE70X-IM/Rev. A0/12-05

TE70X Semi-dry BlottersTE70X Semi-dry transfer unitTE77X Semi-dry transfer unitTE70XP Semi-dry transfer unitTE77XP Semi-dry transfer unit

Page finder Important user information. . . . . . . . . . . . . . . . . . ii

Waste Electrical and Electronic Equipment (WEEE) . . v

1. TE70X Semi-dry blotters: Description . . . . . . . . . . .1

Unpacking . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

Specifications . . . . . . . . . . . . . . . . . . . . . . . . . 3

Important information . . . . . . . . . . . . . . . . . . . 4

2. Operating instructions. . . . . . . . . . . . . . . . . . . . . . .5

3. Care and maintenance . . . . . . . . . . . . . . . . . . . . .14

4. Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . .15

5. Electrotransfer notes. . . . . . . . . . . . . . . . . . . . . . .18

6. Bibliography. . . . . . . . . . . . . . . . . . . . . . . . . . . . .20

7. Customer service information . . . . . . . . . . . . . . . .21

8. Ordering information . . . . . . . . . . . . . . . . . . . . . . .22

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Important user informationPlease read this entire manual to fully understand the safe and effective use of this product. Should you have any comments on this manual, we will be pleased to receive them at:

Hoefer, Inc.953 Indiana StreetSan Francisco, CA 94107 USA [email protected]

Hoefer, Inc. reserves the right to make changes in the specifications without prior notice.

Warranty and liabilityHoefer, Inc. guarantees that the product delivered has been thoroughly tested to ensure that it meets its published specifications. The warranty included in the conditions of delivery is valid only if the product has been installed and used according to the instructions supplied by Hoefer, Inc.

Hoefer, Inc. shall in no event be liable for incidental or consequential damages, including without limitation, lost profits, loss of income, loss of business opportunities, loss of use and other related exposures, however caused, arising from the faulty and incorrect use of the product.

Renseignements importants d’utilizationPour une bonne compréhension et une utilisation en sécurité maximale, il convient de lire entièrement ce manuel. Tous vos commentaires sur ce manuel seront les bienvenus et veuillez les adresser à:


Hoefer, Inc. se réserve le droit d’effectuer des modifications de ces spécifications sans aucun préavis.

Garantie et responsabilitéHoefer, Inc. garantit à l’utilisateur que le produit livré a subi avec succès tous les essais prévus pour s’assurer qu’il est conforme aux spécifications et normes en vigueur. La garantie incluse dans les condi-tions de livraison n’est valable que si le produit a été installé et utilisé conformément aux instructions fournies par Hoefer, Inc.

La société Hoefer, Inc. ne sera en aucun cas responsable de tout dommage causé directement ou indirectement par toute utilisation incorrecte ou non approuvée du produit ou découlant de cette utilisa-tion, y compris toute perte de bénéfice ou de recettes, toute perte de perspectives commerciales, tout empêchement d’utilisation et tout autre risques ayant un rapport avec l’utilisation du produit, mais sans aucune limitation quant à la nature de ces dommages.

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Información importante para el usuarioPara comprender el producto y utilizarlo con seguridad es necesario leer este manual en su totalidad. Si desearan hacer algún comentario sobre este manual, tengan la amabilidad de remitirlo a:


Hoefer, Inc. se reserva el derecho a modificar las especificaciones sin previo aviso.

Garantía y responsabilidadHoefer, Inc. garantiza que el producto entregado ha sido probado a fondo para comprobar el cumplimiento de las especificaciones publica-das. La garantía incluida en las condiciones de entrega sólo es válida si el producto se ha instalado y utilizado de acuerdo con las instrucciones entregadas por Hoefer, Inc.

Hoefer, Inc. no será responsable, bajo ningún concepto, de daños direc-tos o indirectos, incluyendo sin limitación la pérdida de beneficios, la pérdida de ingresos, la pérdida de oportunidades de negocio, la pérdida de utilización y otras consecuencias relacionadas, cualquiera que sea la causa, que se deban a la utilización defectuosa e incorrecta del producto.

Wichtige benutzerinformationenFür ein vollständiges Verständnis und eine sichere Handhabung dieses Produktes ist es notwendig, daß der Benutzer dieses Handbuch voll-ständig durchliest. Wenn Sie Anmerkungen zu diesem Handbuch haben, dann senden Sie diese bitte an:


Hoefer, Inc. behält sich das Recht vor, die Spezifikationen ohne vorhergehende Ankündigung zu ändern.

Gewährleistung and haftungHoefer, Inc. garantiert, daß das gelieferte Produkt sorgfältig auf die Einhaltung der veröffentlichten Spezifikationen getestet wurde. Die in den Lieferbedingungen näher erläuterten Gewährleistungsansprüche gelten nur dann, wenn das Produkt gemäß den von Hoefer, Inc. geliefer-ten Anweisungen installiert und benutzt wurde.

Hoefer, Inc. übernimmt keinerlei Haftung für Schäden oder Folge-schäden, einschließlich, aber nicht begrenzt auf Gewinneinbußen, Einkommensverluste, entgangene Geschäftsabschlüsse, Verlust der Gebrauchsfähigkeit oder andere Verluste, die wie auch immer durch eine fehlerhafte oder unsachgemäße Verwendung des Produkts verursa-cht wurden.

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Informazioni importanti per l’operatorePer un utilizzo sicuro del prodotto, leggere attentamente l’intero conte-nuto del presente manuale. Si prega di inviare eventuali commenti al presente manuale a:


Hoefer, Inc. si riserva il diritto di apportare modifiche ai dati tecnici senza preavviso.

Garanzia e responsabilitáHoefer, Inc. garantisce che prima della consegna il prodotto è stato collaudato a fondo per soddisfare i requisiti specificati. La garanzia inclusa nelle condizioni di consegna risulta valida solamente se il prodotto è stato installato ed utilizzato nel rispetto delle istruzioni fornite da Hoefer, Inc.

Hoefer, Inc. non potrà essere ritenuta responsabile di incidenti o danni consequenziali, inclusi’ma non limitati’a perdite di profitti, mancato guadagno, perdite di affari, difetti di funzionamento e relative espo-sizioni, dovuti ad un utilizzo non corretto del prodotto.

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Waste Electrical and Electronic Equipment (WEEE)

This symbol indicates that the waste of electrical and electronic equipment must not be disposed as unsorted municipal waste and must be collected separately. Please contact an authorized representative of the supplier for information concerning the decommissioning of your equipment.

Ce symbole indique que le gaspillage d’équipement élec-trique et électronique ne doit pas être disposé comme le gaspillage municipal non trié et doit être séparément recueilli. S’il vous plaît contacter un représentant autorisé du fournisseur pour l’information à propos du déclasse-ment de votre équipement.

Dieses Symbol zeigt an, dass die Verschwendung elek-trischer und elektronischer Ausrüstungen als unsortierte städtische Verschwendung nicht verfügt werden muss, und muss getrennt gesammelt werden. Bitte kontaktie-ren Sie einen ermächtigten Vertreter vom Lieferanten für Informationen betreffend des Stilllegens von Ihren Ausrüstungen.

Questo simbolo indica che lo spreco di apparecchiatura elettrica ed elettronica non deve essere disposto lo spreco come unsorted municipale e deve essere separatamente raccolto. Per favore di contattare un rappresentante autorizzato del fornitore per le informazioni riguardanti il rimuovere dal servizio attivo della sua apparecchiatura.

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Denna symbol anger att elektriska och elektroniska utrust-ningar inte får avyttras som osorterat hushållsavfall och måste samlas in separat. Var god kontakta en auktoriserad leverantör representant för information angående avyttring av utrustningen.

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1. TE70X Semi-dry blotters: DescriptionThe TE70X, TE77X, TE70XP, and the TE77XP semi-dry blotters rapidly transfer proteins from polyacrylamide gels onto a membrane by means of a low current and low voltage electrotransfer with minimal Joule heating. Most transfers are complete in one hour or less.

The smaller TE70X unit transfer surface is 14 × 16 cm, suitable for transferring standard gels, including those from the SE 600 Chroma and the SE 400. The larger TE77X transfer surface is 21 × 26 cm, suitable for transferring large format gels.

The TE70X and TE77X have safety circuits built into the instrument that limit the voltage to 30 V and the current to 0.5 A. The circuits protect the user from unnecessary electrical hazards.

The TE70XP and the TE77XP have a built-in power supply for transferring gels. This eliminates the need for an external power supply. These instruments can deliver up to 30 V and 0.5 A. The instruments also monitor the transfer stack resistance, and can stop a transfer if large changes indicate that the buffer system is depleting.

Multiple gels can be transferred at the same time by placing several small gels of the same thickness side by side, or by stacking two gels vertically in a carefully constructed multi-layered stack.

The electrodes are made of the best possible materials to ensure the units last a long time.

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Fig 1. Semi-dry transfer unit main components.

power cords

top electrode

power transformer

lower electrode

TE77XP

voltage leads

top electrode

lower electrode

TE70X

UnpackingUnwrap all packages carefully and compare contents with the packing list, making sure all items arrived. If any part is missing, contact Hoefer, Inc. Inspect all components for damage that may have occurred while the unit was in transit. If any part appears damaged, contact the carrier immediately. Be sure to keep all packing material for damage claims or to use should it become necessary to return the unit.

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The TE70X and TE77X require a power supply with a minimum rating of 0 – 500 mA, 0 – 30 V.

Specifications TE70X TE77X TE70XP TE77XP

Electrode size (cm) 14 × 16 21 × 26 14 × 16 21 × 26

Output: Voltage (V) 30 30 30 30 Current (A) O.5 O.5 O.5 O.5

Inputs: Voltage (V) 100 – 240 100 – 240 Current (A) 0.7 0.7

Frequency (Hz) 47– 63 47– 63

Minimum transfer stack thickness: TE70X, TE70XP 3.2 mm TE77X, TE77XP 2.8 mm

Environmental Indoor use: 4 – 40 °C operating conditions: Humidity up to: 80% Altitude up to: 2000 m Installation category: II Pollution degree: 2

Dimensions (w × d × h): 38 × 46 × 9 cm (15 × 18 × 3.5 in)

Weight: Shipping 6.8 kg Unit 3.7 kg

Certification: EN61010-1, EN 61326, CE UL61010-1-2004 CSA 22.2 61010-1-04

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This declaration of conformity is only valid for the instrument when it is:

• used in laboratory locations.

• used as delivered from Hoefer, Inc. except for alterations described in the user manual.

• connected to other CE labeled instruments or products recommended or approved by Hoefer, Inc.

Important information• The electric components in the transfer unit base

must not become wet. Do not immerse the unit in water. Rinse only the electrodes with distilled water before and after use. (Refer to the Care and mainte-nance section for cleaning instructions.)

• Be sure to use enough buffer-soaked sheets of blot-ting paper on both sides of the membrane/gel stack so that the buffer does not become depleted during the transfer.

• If this equipment is used in a manner not specified by the manufacturer, the protection provided by the equipment may be impaired.

• Only accessories and parts approved or supplied by Hoefer, Inc. may be used for operating, maintaining, and servicing this product.

Informations importantes• Les composants électriques dans la sûreté

enclenchent le logement de la base d’unité de transfert ne doit pas devenir mouillé. Ne pas immerger de partie de l’unité dans l’eau. Rincer seulement les électrodes avec l’eau distillée avant l’usage. (Se référer à la section de Soin et entretien pour nettoyer d’instructions.)

• Etre sûr d’appliquer assez de tampon feuilles trempées de tacher de papier sous la membrane et par-dessus le gel pour empêcher la pile du séchage hors. Les dommages irréparables à l’unité résult-eront si la pile sèche hors et l’unité est permise de surchauffer.

• Si cet équipement est utilisé dans une manière pas spécifié par le fabricant, la protection fournie par l’équipement peut être altérée.

• Seulement les accessoires et les parties ont approuvé ou fourni par Hoefer, Inc. sont recomman-dés pour l’utilisation, l’entretien et réparation de cet appareil.


English

Français

2. Operating instructions To transfer proteins, prepare the unit, assemble the stack, and connect to a power supply if necessary. Then run the transfer for the required amount of time. Each step is described below.

1Prepare the unit by rinsing the electrodes with distilled water.

2Prepare the gel

Cut away the wells and/or stacking gel section. Equilibrate the gel in transfer buffer if required.

3Prepare the transfer stack

Cut the blot paper and transfer membrane to the same size as the gel. Stack the layers carefully so the edges align. If for some reason the membrane needs to be larger than the gel, use a mylar mask (see optional step below) to ensure the current does not bypass the gel.

Optional: Cut a mask to the proper size

Measure the gel. Cut an opening centered in the solid mask to a size roughly 2 mm smaller than the gel. Take care with sharp blades when cutting the masks. Place the mask in the base of the unit, centering the opening.

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Note: Take care to place the gel correctly on the first try because proteins begin to transfer imme-diately; once transfer has begun, moving the gel will distort results or cause “shadow” bands on the blot.

4Prepare the blotting paper

For each gel, cut at least 6 pieces of blotting paper the same size as the gel or slightly smaller.

Gauge the thickness or number of blotting paper layers according to the amount of buffer required; up to 300 ml of buffer may be required for larger gels or for transfers of 60 minutes in order to prevent the stack from drying out or the buffer from becoming depleted.

Saturate at least 3 pieces of the blotting paper with transfer buffer. One by one, center each sheet on the lower electrode and remove all trapped air by rolling a clean pipet or roller from the center toward the edges.

5Prepare the membrane

For each gel, cut 1 membrane the same size as the gel or slightly smaller. (A larger membrane may contact an electrode panel, creating a pathway by which current can bypass the gel.)

Pre-wet nitrocellulose or nylon membranes with distilled water. Pre-wet PVDF or other hydrophobic membranes with methanol. Then soak all membrane types in transfer buffer for 2– 5 minutes.


Note: Each 21 × 26 cm blot paper will absorb approximately 50 ml of transfer buffer. Each 14 × 16 cm blot paper will absorb approxi-mately 20 ml of buffer.

Note: Always wear gloves when handling membranes to avoid leaving fingerprints.

6Complete the stack

a) Place the pre-wet membrane onto the stack of blotting paper.

b) Place the gel on the membrane. Note: Proteins bind to the membrane as soon as contact occurs, so it is important to place the gel correctly on the first try.

c) Cover the gel with three layers of buffer saturated blotting paper.

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Important! Stack each layer with care, with edges parallel. As each layer is added, remove all air pock-ets by rolling a clean pipet from the center to the edges. Add a few drops of buffer to trouble areas to help remove air pockets.

Mask (opening approx. 2 mm smaller than the gel on all sides). Optional.

Blotting paper (2–3 sheets)

Blotting paper (2– 3 sheets)

Gel

Membrane

These stack compo-nents are the same size as the gel or slightly smaller

Fig 2. Transfer stack for a single gel.


Multiple gels: Either lay gels of the same thickness side-by-side (Fig 3), or stack 2 sandwiches layered as shown (Fig 4).

For best results, the transfer stack should be centered in the electrode panels.

If two gels are stacked, separate them with porous cellophane — not plastic wrap! (Cellophane permits electric current to pass but stops proteins.) Cut the cellophane slightly smaller than the gel and wet with transfer buffer.

Several sheets of buffer-soaked blotting paper on each gel provides electrical continuity.

7Place the Cover on top of the transfer stack. Do not remove the cover until after the transfer is complete in order to prevent stack components from moving.

Mask (optional)

Blotting paper, (2–3 sheets)

Blotting paper, (2– 3 sheets)

Gel

Membrane

Note: When transferring multiple gels, transfer efficiency depends on such factors as gel thickness, gel position in the stack, transfer buffer, membrane type, and, most importantly, the characteristics of the protein. The gel closest to the anode generally transfers the most completely. It is preferable to lay gels side-by-side rather than stacking them.

Fig 3. Transfer stacks for gels placed side-by-side.

Note: There must be no electrical contact between the two stacks.

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Blotting paper, (2–3 sheets)

Blotting paper, (1 sheet)

Cellophane

Mask (optional)

Gel

Membrane

Blotting paper, (2– 3 sheets)

Gel

Membrane

Fig 4. Transfer stack for stacked gels.

Separate the stack for each gel with a sheet of cellophane.


Electrotransfer

TE70X and TE77X

1

The power supply should be switched off and both the current and voltage controls set at zero

Then plug the color-coded leads from the base of the transfer unit into the power supply jacks, matching red to red and black to black. Do not reverse polar-ity. The lid contains the black, or negative electrode. The base contains the red, or positve electrode.

2Set the power supply current

The maximum current setting should not exceed 0.8 mA/cm2 of the gel surface. If transferring several layers of gels, a longer transfer time may be required. Use the graph below to quickly find the current setting for your gel size, or calculate the gel area (cm2) and multiply it by 0.8 mA/cm2.

3If available, set the power supply timer

Most transfers are complete within one hour, but larger proteins, proteins from native gels, and thicker gels may require an additional 1 hour of transfer time. The optimum transfer time for each protein and gel system must be determined empirically.

Fig 5. Recommended current settings for different gel sizes.

Note: Generally, smaller fragments transfer more quickly than larger ones.

Note: Transfers exceeding one hour may require additional sheets of buffer saturated blot paper in the transfer stack.

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TE70XP and TE77XPTurn the instrument on using the POWER button on the front keypad.

The instrument works by setting the current and time, and then starting the transfer. During the transfer, the voltage can be displayed. The trans-fer can not be set to run at constant voltage.

The DISPLAY MODE BUTTON toggles between the current (mA), time (hour:minutes) and the voltage (V).

The UP and DOWN arrows to change the value for the current and time.

Press the START/STOP button to start the transfer.

Variable Units Range Increment

Current milliamps 1– 500 1 mA

Time hours:minutes 5 min – 4:00 hours 5 min

Voltage volts 0 – 30 (read only) 1 volt

Note: While setting the time, there is an extra setting labelled “on” above the 4:00 upper limit. This will disable the timer, allowing the transfer to run continuously until manually turned off by the user.

transferstart/stop

up/down arrowsadjust value

display modecycles LED display

power ON/OFF

Fig 6. TE70XP keypad.


During the transfer

TE70XP and the TE77XP

A red LED will appear in the lower right corner of the Display when voltage is applied to the transfer stack.

Both the Time and the Current can be changed as the transfer progresses. Select mA or time using the DISPLAY MODE button, and the UP and DOWN to change the value. While the value is being changed, the LED will blink. After about 10 seconds the LED will stop flashing and show the real time run parameters.

Buffer Depletion

One of the more common failure modes for a Western transfer is buffer depletion. Buffer depletion leads to changes in pH and overheating, both of which are detrimental to the transfer.

The TE70XP and the TE77XP instruments monitor the transfer stack resistance. Large changes in this resistance indicate the depletion of the buffer system. The instrument can stop a transfer before these changes lead to further problems, like burning of the transfer stack.

If this condition happens, the error message “dry” will be on the display. The instrument has stopped the transfer. If desired, the cover can be removed, and buffer can be added to the transfer stack. Replace cover, and press any key (except Power ON/OFF) to continue.

For future transfers, add more layers of buffer soaked blot paper.

Red LED

Dry error message.

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After the transfer is complete

TE70X and TE77X

Turn off the power supply and disconnect the leads from the power supply.

TE70XP and TE77XP

After a transfer is complete, the unit will beep for 5 seconds. The LED display will blink and cycle between the values for the current, time and voltage at the time the transfer was stopped. These can be recorded into a notebook, if desired. Pushing any button will clear the display.

1Remove the cover slowly because the stack may adhere to it.

2Remove and dispose of the upper blot papers.

3Remove the gel(s).

Optional: Stain gel to check for residual protein left in the gel.

4If desired, label the gel contact side of the membrane with a soft pencil.

5Remove the membrane(s) from the stack with blunt forceps. Process the membrane according to your protocol or allow the membrane to air dry prior to storage.

6Remove the remaining blot papers and dispose.

7Rinse the unit according to the Care and maintenance instructions (page 14).

Note: Staining the gel(s) for residual protein gives an indication of the completeness of transfer.

Note: Rewet dried membranes in the appropriate wetting buffer prior to processing.


3. Care and maintenance• Do not autoclave or wash the unit in

a dishwasher.

• Do not immerse the unit in water!

Rinse the cathode in the cover and the anode in the base with distilled water. Let the unit air dry completely. If using radioactive reagents, decontaminate the unit with a cleaning agent such as Contrad™ 70. Never use abrasive cleansers.

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4. Troubleshooting

problem solution

Incomplete transfer

Blank or faint areas on Remove trapped air pockets between the gel and the membrane membrane during stack assembly.

Use buffer with a lower ionic strength.

Molecules do not migrate Check all electrical connections. Confirm that current out of gel is flowing through the transfer stack.

Check that the buffer pH is close to the intended pH. Most buffers should not be titrated. Make fresh buffer.

Use 3.5 mM SDS (0.1%) in the transfer buffer.

Add several more sheets of buffer-saturated blotting paper to each side of the gel sandwich so that more buffer is present during the transfer.

Increase the transfer period. Large fragments may require an additional hour.

Do not use staining or fixing agents on the gel before transfer.

Use a thinner gel.

Reduce the gel acrylamide concentration.

If using a non-nitrocellulose membrane, avoid includ-ing methanol in the transfer buffer or reduce the amount to the minimum possible.

Use reagent-grade chemicals.

Increase the net charge on the protein by using a transfer buffer with a different pH. Lower pH (< 6 – 7) increases the positive charge on proteins; higher pH ( > 6 – 7) increases the negative charge on proteins.


problem solution

Open circuit or no output current Blown fuse in protection circuit. for TE70X or TE77X Replace fuse, (see Accessories in Ordering informa-

tion on page 22).

Smeared or diffuse band patterns If equilibrating before the transfer, shorten or elimi-nate the equilibration time and/or equilibrate under coldroom conditions.

If the transfer buffer contains ≥10% methanol, equili-brate the gel in transfer buffer for 30 minutes to allow it to shrink before assembling the stack. Note: Large proteins may not migrate as readily once the pore size is slightly reduced.

Take care that the gel does not shift once it contacts the membrane.

Buffer depletion can change the pH in the transfer stack, and have a negative effect on the transfer. On subsequent transfers, either shorten the transfer time, reduce the current, or increase the number of buffer-soaked blotting papers in the stack.

Check that the preferred binding surface of the membrane (if any) contacts the gel.

Uneven band transfer The blotting paper and membrane must be the same size as the gel or 1– 2 mm smaller. Larger sizes will provide an electrical path for current to bypass the gel solution.

Different proteins will transfer at different rates depending on size and net charge.

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problem solution

Inefficient binding to membrane

Chemical parameters Prepare transfer buffer without SDS. (SDS can improve transfer efficiency but can interfere with protein binding to a PVDF membrane.)

Add 10 – 20% methanol to the transfer buffer to enhance binding to nitrocellulose or PVDF.

Membrane parameters Use a membrane with a smaller pore size (0.20 µm) if proteins pass through the membrane.

Place a membrane both over and under the gel to capture any proteins migrating in the opposite direction.

Check if too much sample is available for the binding surface area by placing two membranes instead of one. If “blow through” occurs, reduce the sample load.

Wear gloves when handling membranes.

Store membranes at ambient temperature and out of direct sunlight.

Check shelf life of nitrocellulose membrane, replace if necessary.

TE70XP, TE77XP

Current reading Maximum voltage (30 V) has been reached. lower than setting Buffer may be depleted.

No output current Transfer stack is not making contact with the upper electrode. Add buffer soaked blot papers.

“dry” error message Instrument has detected large changes in transfer stack resistance.

Add more buffer soaked blot papers in future transfers.

No output voltage Transfer stack is being short circuited.

Note: For more troubleshooting ideas, refer to Bjerrum, O.J. et al. (1988).


5. Electrotransfer notes• Run the transfer as soon as possible after

electrophoresis to minimize protein diffusion within the gel.

• Stacked gels must all be the same size.

• Limit transfers to two hours or less.

• The recommended methanol concentration for different membrane types are:

membrane type methanol %

Charged nylon 0

Nitrocellulose 10 – 20

PVDF 10 – 20

• Use a buffer with low ionic strength such as one of the two listed below to prevent over-heating. Use the CAPS buffer when Tris cannot be used (e.g., peptide sequencing). CAPS can improve transfer because of its effect on the charge of the protein (see Matsudaira, 1987).

Towbin buffer(25 mM Tris, 192 mM glycine, 20% v/v methanol, pH 8.3, 1 liter)

Tris (FW 121.1) 25 mM 3.0 g

Glycine (FW 75.07) 192 mM 14.4 g

SDS* (FW 288.4) 0.1% (3.5 mM) 1.0 g

Dissolve in 600 ml distilled water. Add methanol as required†. Bring to 1 liter with distilled water. Do not adjust the pH, which should be between 8.2 – 8.4. Optional: Chill before use.

*Optional: Adding SDS can improve transfer efficiency.† Depending on the membrane type selected (see table above), adding methanol can improve transfer results.

Note: Buffers containing methanol may deteriorate if stored for long periods — add methanol just prior to transfer.

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CAPS buffer, 1X(10 mM CAPS, pH 11.0, 1 liter)

CAPS (FW 221.3) 10 mM 2.2 g [3-(cyclohexylamino)-1-propanesulfonic acid]

Dissolve in 600 ml distilled water, adjust to pH 11.0 with conc. NaOH. Adjust volume to 1.0 liter.

• For a 3-buffer system, refer to Kyhse-

Anderson, J. (1984).

• Transfer efficiency varies depending on the gel concentration, which can be optimized. For more information, refer to Smejkal and Gallagher (1994).


6. BibliographyBjerrum, O.J., Larsen, K., and Heegaard, N., CRC

Handbook of Immunoblotting of Proteins 1, Section 7. CRC Press (1988).

Gallagher, S., Winston, S.E., Fuller, S.A. and Hurrell, J.G.R., Immunoblotting and Immunodetection. In Current Protocols in Molecular Biology. 10.8.1–10.8.17. Greene Publishing and Wiley-Interscience, NY (1993).

Hancock, K. and Tsang, V., India ink staining of proteins on nitrocellulose paper. Anal. Biochem. 133, 157–162 (1983).

Kyhse-Anderson, J., Electroblotting of multiple gels: A simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitro-cellulose. J. Biochem. and Biophys. Meth. 10, 203–209 (1984).

Matsudaira, P., Sequence from Picomole Quantities of Proteins Electroblotted onto Polyvinylidene Difluoride Membranes. J. Biol. Chem. 262, 10035 (1987).

Sasse, J. and Gallagher, S., Detection of Proteins on Blot Transfer Membranes. In Current Protocols in Molecular Biology. 10.7.1–10.7.3. Greene Publish-ing and Wiley-Interscience, NY (1991).

Smejkal, G., and Gallagher, S., Determination of Semidry Protein Transfer Efficiency with Transverse Gradient Gel Electrophoresis. Biotechniques. 16, 196–202 (1994).

Tovey, E. and Baldo, B., Comparison of semi-dry and conventional tank-buffer electrotransfer of proteins from polyacrylamide gels to nitrocellulose

membranes. Electrophoresis 8, 384–387 (1987).

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7. Customer service information

Technical service and repairHoefer, Inc. offers complete technical support for all our products. If you have any questions about how to use this product, or would like to arrange to repair it, please call or fax your local Hoefer, Inc. representative.

Check the Hoefer, Inc. website for the distributor in you area.

www.hoeferinc.com

Important! Request a copy of the Hoefer, Inc. “Health and Safety Declaration” form before return-ing the item. No items can be accepted for servicing or return unless this form is properly completed.


8. Ordering information

product qty. order no.

TE70X Semi-dry Transfer Unit, 14 × 16 cm 1 TE70X Includes 25 sheets of blotting paper, 50 sheets of

cellophane, 2 solid masks, masks for 7 × 8 cm and for 14 × 16 cm gels

TE77X Semi-dry Transfer Unit, 21 × 26 cm 1 TE77X Includes 25 sheets of blotting paper, 50 sheets of

cellophane, 2 solid masks, mask for 14 × 16 cm gels

TE70XP Semi-dry Transfer Unit, 14 × 16 cm 1 TE70XP Includes 25 sheets of blotting paper, 50 sheets of

cellophane, 2 solid masks, masks for 7 × 8 cm and for 14 × 16 cm gels

TE77XP Semi-dry Transfer Unit, 21 × 26 cm 1 TE77XP Includes 25 sheets of blotting paper, 50 sheets of

cellophane, 2 solid masks, mask for 14 × 16 cm gels

Accessories

TE70X and TE70XP

Solid masks, 16.5 × 18.5 cm 4 TE74

Porous cellophane, 20 × 35.5 cm 50 TE73

Blotting paper, precut, 14 × 16 cm 25 TE76-1416

TE77X and TE77XP

Solid masks, 23 × 27.5 cm 4 TE78

Porous cellophane, 35 × 44 cm 50 SE1142

Blotting paper, precut, 21 × 26 cm 25 TE76

Power Cord Kit 1 set PS36-24

TE70X Semi-dry blotters User Manual (this manual) 1 TE70X-IMA0

Fuse 1.6 A, 250 V, SB 5 × 20 5 PSF1.6A-MICRO

• p23

Related products

product order no.

SE600 Chroma SE600X-15-1.5

MiniVE Vertical Electrophoresis system SE300-10A-1.0

PS 2A200 Power Supply PS2A200

Transfer Membranes

Pure Nitrocellulose, sheets and roll

0.45 µm pore size

8 × 9.5 cm, 10 sheets GM-NC45-89

16 × 16 cm, 10 sheets GM-NC45-1616

20 × 23.5 cm, 10 sheets GM-NC45-2320

30 cm × 3 m, 1 roll GM-NC45

0.2 µm pore size

30 cm × 3 m, 1 roll GM-NC22

PVDF membrane

0.45 µm pore size

30 cm × 3 m, 1 roll GM-PV45

Caliber Electrophoresis Reagents

Tris, 1 kg GR132-1

Glycine, 1 kg GR125-1

SDS, 500 g GR126-500

Tween® 20, 500 ml GR128-500

DTT, 5 g GR122-5

Glycerol, 1 L GR124-1

Bromophenol Blue (BPB), 10 g GR120-10

Protein Determination Reagent, 500 standard assays GR133-500

Coomassie® Brilliant Blue G-250, 25 g GR134-25

Coomassie Brilliant Blue R-250, 25 g GR135-25


Notes:

• p25

Notes:


Notes:


Hoefer, Inc. 953 Indiana Street San Francisco, CA 94107 USA

www.hoeferinc.com

© 2005 Hoefer, Inc. — All rights reserved.

Printed in the USA

Caliber, Chroma and MiniVE are trademarks of Hoefer, Inc.

Contrad 70 is a trademark of Decon Laboratories.

Tween is a trademark of ICI Americas Inc.

Coomassie is a trademark of ICI plc.

te70x semi-dry blotters · 2012. 10. 17. · tions de livraison n’est valable que si le produit a...

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